Alkaline preparaions of Inula for the control of fungal diseases in plants

ABSTRACT

Rapid, convenient and inexpensive methods for preparing fungicidal suspensions from Inula are described. Methods for controlling fungal disease and crops by using alkaline solutions are also disclosed.

[0001] This application claims priority of U.S. Provisional ApplicationSerial No. 60/072,356 filed Jan. 23, 1998.

FIELD OF THE INVENTION

[0002] The subject invention relates an anti-fungal preparation for thecontrol of fungal diseases in plants, more specifically, to alkalineextracts of Inula which are highly active in controlling diseases causedby fungi in crop plants.

BACKGROUND OF THE INVENTION

[0003] Extracts of plants which are members of the Inula species areeffective against infections of plants caused by a variety of fungi.These extracts are typically prepared by dipping freshly cut Inulashoots in an organic solvent or by agitating freshly cut or dried Inulashoots in an organic solvent, removing the solvent to form a paste, andthen dissolving the paste in an organic solvent or in water, possiblywith an additive. Alternatively, suspensions of Inula can be prepared bygrinding dried Inula shoots into a fine powder, adding an emulsifier,and then suspending the mixture in water. In both cases, the resultantpreparation is then applied to plants resulting in the control of a widevariety of fungal diseases. Extracts of suspensions of Inula plants areeffective at low concentrations, in the range of fractions of a singlepercent of extract, such that dilute concentrations have excellentfungal-control properties.

[0004] The plants which are the basis of these fungicidal extracts andsuspensions are Inula viscosa and Inula graveolens (Family Compositae),perennial weeds widespread in the Mediterranean Basin.

[0005] Methods of preparing aqueous extracts from various parts of theInula plant are well known in the literature. Additionally, organicextracts of the Inula plant are also well known in the art, e.g., U.S.Pat. No. 5,837,253. However, the prior art indicates that aqueousextracts of Inula shoots are poorly effective against fungal diseases ofcrop plants whereas extracts made with organic solvents are highlyeffective anti-fungal agents.

[0006] Methods have also been described for using organic solvents toextract Inula plants, but these are clearly distinguished from thatdisclosed in U.S. Pat. No. 5,837,253. Two of the prior art methodsinvolved contacting the whole Inula plant, or the aerial parts thereof,with an organic solvent either by maceration of the plant in the organicsolvent or by percolation of the solvent through the plant. Furthermore,U.S. Pat. No. 4,254,112 to Debat et al., (hereinafter referred to as“Debat”) describes the preparation of extracts of Inula viscosa andInula graveolens using whole Inula plants which have been dried andground and organic solvents, by using a Soxhlet apparatus. The yield ofthe paste obtained by this method was approximately 1.75-4%. U.S. Pat.No. 5,176,193 to Honerlagen et al. (hereinafter referred to as“Honerlagen”) describes a process for preparing a partial extract fromroots of Inula helenium which involves contacting the plant materialwith an organic solvent, adding a drying agent to the solution to removethe water, removing the drying agent, and then distilling the driedorganic phase. By contrast, the method disclosed in U.S. Pat. No.5,837,753 disclosed either briefly dipping the leaves and stems of theshoots of Inula viscosa or Inula graveolens into an organic solvent orshaking the freshly cut or dried and ground leaves and stems of theshoots in an organic solvent for thirty minutes, and then evaporatingthe solvent to form a paste. The yield obtained by this method can be asmuch as 30%, in contrast to the low yields known in the literature anddescribed above.

[0007] The medicinal properties of Inula extracts in humans are wellknown. For example, Debat disclosed the anti-microbial activity ofextracts of Inula for use in human beings. However, the fungicidaleffects of Inula extracts have only been demonstrated on fungi growingin Petri dishes or on post-harvest of fruits. For example, Qasem et al.(Phytopathologia Mediterana, 34:7-14, 1995) demonstrated that the growthof certain fungi in Petri dishes was inhibited by aqueous extracts ofInula viscosa as well as by dried plant material added directly to thefungal growth media. By contrast, the method of the present inventionuses Inula extracts prepared with alkaline aqueous solvents used againstfungal infections of crop plants themselves.

[0008] Clearly, although Inula extracts have been shown to havefungicidal activity in the Petri dish (in vitro) and on plant (in vivo),the methods of preparation for these extracts have not been sufficientfor large scale use directly on crop plants and have had other seriousdisadvantages. The true effectiveness of these extracts against fungalinfections of plants is, therefore, unknown. Furthermore, there is aclear need for better methods to prepare Inula extracts. Qasem et al.(Ibid, page 13, 1995) concluded: “The diversity and the methodology ofextraction and the differences in the results obtained . . . increasedthe need for developing more efficient, convenient, and cheaper methodsof extraction to facilitate more extensive utilization of fungicidalextracts, especially if greater quantity of extracts must be preparedfor large-scale production”.

[0009] Thus, there is a widely recognized need for and would be highlyadvantageous to have a method or methods for preparing extracts and/orsuspensions of plants of Inula species which would facilitate thelarge-scale use of these extracts and suspensions which would simplifytheir preparation and use, be highly effective in controlling fungalinfection in plants, as well as controlling fungal infections in cropplants.

SUMMARY OF THE INVENTION

[0010] According to the present invention, there is provided a methodfor preparing an aqueous extract from Inula species which comprisescontacting shoots and/or portions thereof with an alkaline aqueoussolvent to form a solution and debris, and removing the debris from thesolution.

[0011] There is also provided a method for protecting plants againstfungal infection comprising preparing a fungicidal extract of Inulaspecies by contacting shoots and/or portions thereof with an alkalineaqueous solvent to form an extract solution and debris, and applying afungicidally effective amount of the fungicidal extract to a plant forprotecting the plant against fungal infection.

[0012] There is also provided in accordance with the present invention,a method for preparing a fungicide derived from Inula species whichcomprises combining a substantially powdered form of Inula shoots and/orportions thereof with a solid chemical to form a mixture which upondissolution of the mixture in an aqueous solvent, forms an alkalineaqueous solution which can be directly applied to plants to preventfungal infection.

[0013] Additionally, also in accordance with the present invention,there is provided an alkaline anti-fungal composition which comprisesInula shoots and/or portions thereof and a compound which when mixedwith an aqueous solvent forms an alkaline solution, whereby thecomposition can be directly applied to plants to prevent fungalinfection.

[0014] Accordingly, the subject invention provides a method forunexpectedly producing an anti-microbial extract from Inula plants whichis not only active in vitro against fungi but also is active againstdiseases caused by fungi in crop plants while being safe to apply to thecrop itself.

DETAILED DESCRIPTION OF THE INVENTION

[0015] The present invention provides a method for preparing an extractfrom Inula species including the steps of contacting shoots and/orportions thereof with an alkaline aqueous solvent to form a solution anddebris and then removing the debris from the solution.

[0016] Preferably, the portion and/or portions of the Inula plant whichare utilized in the present invention are the leaves and stem of theupper, younger parts of the shoots, and the upper part of the shootextending from about 20-40 centimeters from the tip of the shoot. Theflowers themselves are not used. As used herein, the term “shoot” isused to define the leaves and stem of the shoot of the Inula plant. Inthe methods of the present invention, the shoots or portions thereof canbe either freshly cut or may be dried prior to the addition of thealkaline aqueous solvent.

[0017] The alkaline aqueous solvent can include any compound orcompounds which alone or in solution have a pH which is greater thanabout seven such as an inorganic base or salt thereof.

[0018] Preferably, the alkaline aqueous solvent includes at least one ofamino compound. The amino compound can include an ammonium compound,organic amines and/or heterocyclic amines. The organic amines caninclude compounds such as ethylamine, ethylenediamine, diaminopropane,diethanolamine, triethanolamine, and butylamine. Heterocyclic compoundscan include compounds such as piperazine and piperidine.

[0019] The removal of the debris from the solution can be accomplishedby filtering the debris from the solution utilizing a screen such as a100 micron μm (micron) sieve or other suitable filtering devices knownto those skilled in the art.

[0020] If dried shoots and/or portions thereof are utilized in themethod of the present invention, the dried shoots and/or portionsthereof can be ground, by various means known to those skilled in theart, to form a powder.

[0021] In an embodiment of the invention, fresh Inula shoots are dippedin the alkaline solution for an appropriate period of time, typicallyfor approximately 0.2-5 minutes. The plant material is then discardedand the solution, which may be colored, obtained therefrom is screenedthrough a filter such as a 100 micron sieve and can be applied, as is,for spraying crop plants.

[0022] In another embodiment, fresh shoots are first air dried at 30-40°C. for period of time sufficient to dry the shoots, typicallyapproximately 1-2 days and the dried shoots then dipped in the alkalinesolution for a period of time which can be approximately 1-10 minutes.The plant material is discarded and the solution (colored) obtained canbe filtered through a filter or sieve and then utilized, as is, forspraying crop plants.

[0023] In another embodiment, dried shoots obtained by air drying asdescribed immediately above can be ground and then passed through ametal sieve (i.e., 2 mm). The ground plant material can then be shakenin an alkaline solution for 1-10 minutes, screened through a filteringdevice such as a 100 micron sieve, and the colored liquid obtainedtherefrom can be utilized for spraying crop plants.

[0024] In yet another embodiment, dried and ground plant material asimmediately described above, can be placed in a mortar and is milled toless than 100 micron-sized particles to obtain a fine powder. The powderis then mixed with an alkaline solution and can be sprayed, as such,onto the crop plants.

[0025] In another embodiment, dried powder which is ground to less than100 micron-sized particles is milled with crystallized cellulose (AvicelPH-101, Fluka) and is then mixed with the alkaline solution and sprayeddirectly onto crop plants.

[0026] In another embodiment, a dried powder which has been milled toless than 100 micron-sized particles is milled with a surfactant such asVercoryl-S, diatomaceous earth, or sodium dodecyl sulfate and is thenmixed with the alkaline solution and is sprayed directly onto the cropplants.

[0027] In another embodiment, dried Inula shoots ground to less then 100micron-sized particles are mixed with a solid chemical, which upondissolution in water, produces an alkaline solution. The solid chemicalcan be any of the compounds described above for the alkaline solutionincluding an imidazole, dibasic ammonium phosphate, Trizma-base (Sigma)either with or without the addition of an inert surfactant formulation.The powder mixture is then mixed with water to form an anti-fungal agentand is sprayed directly onto crop plants. In this form, the anti-fungalagent can be prepared and/or sold as a dry mix which, when hydrated,forms an anti-fungal composition which can be easily applied to plants.In this form, the costs of shipping and/or transporting the anti-fungalmaterial can be reduced since the water would only be added just priorto use/application to plants.

[0028] In another embodiment, the dried and ground Inula of particlesize less than 100 microns can be mixed with (NH4)₂HPO₄ (dibasicammonium phosphate) and be suspended in water to which NH₄OH is added.This solution can then be sprayed directly onto crop plants.

[0029] According to the present invention, there is also provided amethod for protecting plants against fungal infections by the steps of(a) preparing a fungicidal extract of Inula species by contacting shootsand/or portions thereof with an alkaline aqueous solvent to form anextract solution and debris and (b) applying a fungicidally effectiveamount of the fungicidal extract to a plant for protecting a plantagainst fungal infection. As defined herein, a fungicidally effectiveamount includes such an amount that is capable of preventing,inhibiting, and/or killing fungus in or on a plant.

[0030] Solutions or emulsions containing the Inuta extract, or the Inulasuspension, are sprayed onto the upper leaf surfaces of various cropplants by well known techniques for the control of fungal plantinfections. The Inula extracts of the present invention can be used tocontrol fungal plant infections caused by fungi including, but notlimited to, Oomycetes, Ascomycetes, Basidiomycetes, and Fungi imperfecticlasses.

[0031] The method and fungicidal extracts of the present invention areparticularly suitable for use against fungi of the Oomycetes,Ascomycetes, Basidiomycetes and Fungi imperfecti classes including butnot limited to Phytophthora infestans, Pseudoperonospora cubensis,Plasmopara viticola, Sphaerotheca fuliginea, Cladosporium cucumerinum,Erysiphe graminis, Uromyces appendiculatus, and Botrytis cinera and incrops including, but not limited to, grapevines, tomato, wheat, barley,tobacco, potatoes, onions, cucurbits, beans and crucifers.

[0032] Also according to the present invention, there is provided amethod for preparing a fungicide derived from Inula species whichincludes combining a substantially powdered form of Inula shoots and/orportions thereof with a solid chemical to form a mixture which upondissolution of the mixture and an aqueous solution, forms an alkalineaqueous solution which can be directly applied to plants to preventfungal infection. Fungicides prepared by this method are described aboveand in the Example section below.

[0033] Also in accordance with the present invention, an alkalineanti-fungal composition of Inula shoots and/or portions thereof and acompound which when mixed with an aqueous solvent forms an alkalinesolution which can be directly applied to plants to prevent fungalinfection is disclosed. The compounds which can be mixed with an aqueoussolution to form an alkaline solution are described above and in theExamples section below.

[0034] The invention of the present invention is illustrated below inthe following Examples which describe the preparation and use of Inulaextracts and suspensions against fungal infections in plants.

[0035] To test the efficacy of these extracts and suspensions,experiments were performed in which plants were sprayed using a fineglass atomizer with either Inula extract or with pure solvent as acontrol for the Inula extracts, with either Inula extract and emulsifieror with an emulsifier solution as a control for the emulsified Inulaextracts, and with either Inula suspension or with water or emulsifiersolution as a control for the Inula suspensions. Treated and controlplants were then inoculated with a crop-respective fungal pathogen.After an incubation period, the extent of the infection was measured.Unless otherwise stated, percentage protection from the disease due tothe treatment with the Inula extract or suspension was calculated as:

% control of the disease=[1-(% infection and treated plants/% infectionin control plants)]×100.

[0036] The following experimental data illustrate the utility of theflungicidal compositions of the present invention.

EXAMPLES

[0037] METHODS: The crop plants and pathogens used for inoculation aredescribed in the following list. crop plant pathogen disease TomatoPhytophthora infestans late blight Potato ″ ″ Cucumber Pseudoperonosporacubensis downy mildew Melon ″ ″ Wheat Erysiphe graminis tritici powderymildew Cucumber Sphaerotheca fuliginea ″ Melon ″ ″ Tobacco Perenosporatabacina downy mildew Grapes Plasmpara viticola ″ Tomato Botrytiscinerea gray mold Cucumber ″ ″ Tomato Cladosporium fulvum leaf mold #done by blowing spores over the treated (and control) plants. Inoculatedplants were then kept in a growth chamber as above.

Example 1

[0038] Activity of dipping extract of fresh Inula shoots in 0.1 N NaOHfor ten minutes on disease development in crop plants. % Diseased leafarea powdery mildew Extract late blight in potato in wheat controluntreated 94 63 original (pH 12.7) 15 24 diluted: 2 9 25 diluted: 4 1030 diluted: 8 8 50 original (neutralized to pH 7.2) 5 12 diluted: 2 9418 diluted: 4 95 25 diluted: 8 95 30

Example 2

[0039] Activity of dipping extracts of fresh Inula shoots in ammoniumhydroxide (containing 25% NH₃ by weight) for ten minutes on late blightdevelopment in potato. Treatment Diseased leaf area % Protectionuntreated control 98 — NH₄OH 8% 100 0 (=2% aqueous NH₃) 4% 100 0 2% 1000 1% 100 0 Inula in NH₄OH 8% 10 90 4% 20 80 2% 50 50 1% 88 12 # % from aliquid containing 250 g NH₃ per liter. For example, 8% NH₄OH means 8 mlof a solution (containing 25% NH₃) per 100 ml water. To express % NH₄OHas % NH₃, should divide by 4.

Example 3

[0040] Efficacy of Inula extracts, made by dipping fresh shoots in NH₄OHfor ten minutes, in the control of late blight in potato (a) and powderymildew in wheat (b). % blighted leaf area 3d - old extract NH₄OH conc.,% fresh extract 1d - old extract a b untreated control 100 100 100 75 123 63 25 23 2 5 13 23 1 4 8 18 8 11 8 8 10 25 24 16 3 10 5 6

Example 4

[0041] Activity, after storage, of fresh shoot Inula extract made withammonium hydroxide against fungal diseases of crop plants. % diseasedleaf area downy powdery downy storage late blight mildew in mildewmildew in NH₄OH period, in potato, melon in wheat grapes conc., % days 5d 6 d 7 d 10 d control 1 98 81 100 50 untreated 0.25 38 15 50 20 0.5 154 13 15 1.0 8 1 1 5 control 9 98 50 untreated 0.25 69 38 0.5 13 13 1.013 nd control 18 75 untreated 0.25 nd 0.5 0 1.0 0 0 60 95 0.25 38 0.5 211 18

Example 5

[0042] Activity of aqueous or acetone extracts made from dry crushedshoots of Inula against fungal diseases of crop plants. % Diseased leafarea downy powdery late blight mildew in mildew in powdery in potatocucumber cucumber mildew in solvent 4d 6d 6d wheat, 8d untreated control83 88 88 88 water 18 5 75 63 KOH 0.1N 88 4 75 63 NaOH 0.1N 63 28 50 25NH₄OH 1.4%  1 8 50 13 NH₄OH 7%  3 4 25 15 (NH₄)₂HPO₄  0.3 0 50 3 0.1M(1.3%) K₂HPO₄ 0.1M 23* 3 88 20 Acetone  8 3 25 5

Example 6

[0043] Activity of dry crushed Inula shoots extracted with ammoniumhydroxide and ammonium phosphate (dibasic) against fungal disease incrop plants. % diseased leaf area pow- pow- dery dery late late downymildew mildew blight in blight in mildew in in in tomato potato cucumberwheat melon* solvent, conc. pH 4 d 6 d 8 d 11 d 14 d None — 88 100 100100 180 (NH₄)₂HPO₄ 7.9 25 14 16 18 150 1.32% (0.1M) NH₄OH 0.7% 10.97 2522 28 15 55 (0.1M) (NH₄)₂HPO₂ + NH₄OH, mixed, 0.1M   9 + 1 8.25 29 5 2715 80 7.5 + 2.5 8.6 31 13 16 3 100   5 + 5 9.0 11 6 6 0 40 2.5 + 7.5 9.420 0 11 1 25   1 + 9 9.8 16 2 16 8 20 water 6.2 75 80 58 75 180 acetone— 0 16 11 8 130

Example 7

[0044] Activity of crushed dry Inula shoots extracts made with aqueoussolutions of ethylamine and ethylenediamine against fungal diseases ofcrop plants. Potato late blight cucumber downy solvent, conc. and 4dmildew 6d pH % blighted leaf area lesions/plant control untreated 94 125ethylamine 0.7% 15 5 (pH 12.3) ethylenediamine 0.6% 50 4 (pH 11.95)

Example 8

[0045] Activity of alkaline extracts or acetone of fresh Inula shootsagainst disease development in crop plants. % infected leaf area lateblight in tomato powdery mildews in solvent and conc. 6d wheat 11duntreated control 88 75 ethylamine, 1% 0 38 ethylenediamine, 0.9% 0 38ammonium hydroxide, 1% 1 20 (0.25% NH₃ in water)2-amino-2-methylpropanole 25 nt (AMP) 0.9% acetone 0 23

Example 9

[0046] Sequential extraction of fresh Inula shoots with acetone andammonium hydroxide and the activity of the extract against plantdisease. % infected leaf area powdery late blight in mildews in Firstsolvent Second solvent tomato, 6d wheat, 11d untreated control 88 75NH₄OH, 1% — 1 20 acetone — 0 23 NH₄OH, 1% acetone 1 25 acetone NH₄OH, 1%38 75

Example 10

[0047] Antimicrobial activity, in vitro, of alkaline extracts of freshInula shoots. fungal colonies bacterial colonies extraction made withper dish per dish no extract, control 75 30 ethylamine, 1% 7 30 ethylenediamine, 0.9% 10 40 ammonium hydroxide, 1% 12 15 acetone (positivecontrol) 4 0

Example 11

[0048] Dose - dependent activity of alkaline extracts made from drycrushed Inula shoots with 1:1 mixture (pH 9.0) of 0.1M (1.32%)(NH₄)₂HPO₄ and 0.1M (0.7%) NH₄OH, against late blight in tomato. g dryshoots blighted leaf area per 50 ml solvent %, 4 days untreated control88 solvent control 88 0.1 75 0.2 63 0.4 25 0.6 25 0.8 18 1   14 2    3

Example 12

[0049] Activity of aqueous alkaline extracts made from dry crushed Inulashoots against late blight in tomato. % diseased leaf area solvent Inulaextract solvent control untreated control 81 — piperazine hydrate O.05M3 81 (l%) pH 113 piperazine hydrate 0.1M 3 81 (2%) pH 11.3triethanolamine 0.1M 23 81 (1.59%) pH 10.3

Example 13

[0050] The effect of concentration of piperazine hydrate on the activityof Inula extracts against late blight in tomato and potato. piperazinehydrate % diseased leaf area conc.,% tomato potato 0 91 95 0.06 18 250.12 11 25 0.25 4 25 0.50 18 44 1.00 0 75 2.00 8 75 (=0.1 M)

Example 14

[0051] Efficacy of tetraethylene or piperidine aqueous solutions onextraction of antifungal components from dry shoots of Inula % diseasedleaf area Tomato late cucumber downy wheat powdery Treatment blight, 4dmildew, 6d mildew, 10d none (control) 88 88 50 tetraethylene 1% 63 31 50(pH 11.2) piperidine 1% 88 94 25 (pH 12.0) Inula in tetraethylene 25 4750 1% Inula in piperidine 1% 25  1 13

Example 15

[0052] Activity of aqueous alkaline or acetone extracts made from Inulashoots against mildews in cucumber and wheat % diseased leaf area downymildew powdery mildew powdery mildew in cucumber in cucumber in wheatsolvent 6d 11d 8d untreated control 75 100 100 Trizma-base ® 20 13 3(Sigma) 1% piperazine 18 38 38 hydrate 1% piperadine 1% 5 25 18imidazole 1% 0 0 0 acetone 2 25 25 (positive control)

Example 16

[0053] Efficacy of sec - butylamine in extracting antifungal componentsfrom dry Inula shoots. lesions per plant % diseased leaf area sec -butylamine late blight in late blight powdery mildew conc., % potato 3dtomato 4d cucumber 13d wheat 7d 0 130 88 88 100 0.06 60 63 14 23 0.12 025 14 0 0.25 5 8 9 15 0.5 0 15 15 3 1.0 (pH 11.5) 2 3 9 5

Example 17

[0054] Efficacy of imidazole in extracting antifungal component from dryInula shoots. % diseased leaf area late blight late blight powderymildew Treatment tomato, 5d potato, 5d wheat, 8d none 81 100 69Imidazole only 0.12% pH 9.55 95 50 63 0.25% pH 9.70 88 50 50  0.5% pH9.86 69 25 23   1% pH 9.95 50 18 25 Inula extract in imidazole 0.12% 305 0 0.25% 20 18 18  0.5% 30 30 0  1.0% 30 15 0 Inula in acetone 5 0 3

Example 18

[0055] Composition of a 50 WP formulation of Inula shoots and itsactivity against fungal plant diseases. Composition:   5 g dry Inulashoot powder 2.5 g imidazole 2.5 g inert carriers and surfactants(Sandoz) Activity % diseased leaf area g product per late blight lateblight powdery mildew 50 ml water pH tomato 5d potato 5d wheat 13d 0 —98 100 38 0.25 8.20 75 75 25 0.5 8.21 38 63 13 0.75 8.22 31 25 0 1 8.2323 18 0 1.25 8.24 20 20 0 1.5 8.25 15 13 0 2 8.25 1 10 0

Example 19

[0056] Composition of a 50 WP formulation of Inula shoots and itsactivity against fungal plant diseases. Composition:   5 g dry Inulashoot powder 3.75 g imidazole 1.25 g inert carriers and surfactants(Sandoz) Activity % diseased leaf area g product per late blight lateblight powdery mildew 50 ml water pH tomato 4d potato 5d wheat 9d 0 — 9581 38 0.25 8.28 75 69 25 0.5 8.29 63 25 0 0.75 8.32 38 15 0 1 8.33 25 130 1.25 8.35 21 13 0 1.5 8.36 13 18 0 2 8.37 6 8 0

Example 20

[0057] Composition of 45 WP formulation of Inula shoots and its activityagainst fungal plant diseases. Composition: 5 g dry Inula shoot powder 5g imidazole 1 g inert carriers and surfactants (Sandoz) Activity %diseased leaf area g product per 50 ml late blight in water pH tomatoes4d 0 — 75 0.25 8.18 75 0.5 8.23 50 0.75 8.28 50 1 8.29 38 1.5 8.30 25 28.38  3

Example 21

[0058] Composition of 50 WP formulation of Inula shoots and its activityagainst fungal plant diseases. Composition:   5 g dry Inula shoot powder2.5 g dibasic ammonoim phosphate 2.5 g Avicel PH - 101 ® (Fluka) =crystalline cellulose % diseased leaf area Activity downy g product lateblight mildew powdery per 50 ml late blight in in potato, in cucumber,mildew in water pH tomato, 4 d 6 d 6 d wheat, 9 d 0   — 98 98 50 75 0.17.33 75 93 13 75 0.2 7.44 75 93 8 50 0.4 7.46 63 38 3 50 0.6 7.46 63 301 25 0.8 7.46 18 23 3 23 1   7.46 23 20 0 20 1.5 7.46 23 18 0 18

Example 22

[0059] Composition of 50 WP formulation of Inula shoots and its activityagainst fungal plant diseases. Composition:   5 g dry Inula shoot powder2.5 g Trizma - Base ® (Sigma) 2.5 g Avicel PH - 101 ® (Fluka) Activity %diseased leaf area g product per late blight late blight in powderymildew 50 ml water pH tomato, 3d potato, 5d in wheat, 8d 0 — 95 95 750.2 8.55 88 95 50 0.4 8.62 88 93 50 0.6 8.67 38 69 25 0.8 8.68 23 38 251 8.70 30 25 25 1.5 8.72 23 25 25 2 8.74 25 25 25

Example 23

[0060] Composition of 50 WP formulation of Inula shoots and its activityagainst fungal plant diseases. Composition: 5 g dry Inula shoot powder 5g dibasic ammonium phosphate Activity % diseased leaf area g productlate blight in potato per 50 ml late blight in lesions/plant powderymildew water pH tomato, 4 d 3 d 4 d in wheat, 7 d 0   — 95 >500 88 880.2 7.42 38 200 69 38 0.4 7.45 25 180 69 38 0.6 7.45 23 50 25 23 0.87.45 20 40 25 13 1   7.46  3 20 20 13 2   7.47  1 5  8  3

Example 24

[0061] EXAMPLE 24 Composition of alkaline wettable powder formulationsof Inula used to control plant diseases. Percent Other Formula a-iAlkaline agent pH components Inula leaves (27%) 25 none 6.8 Zeofree 80(60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%) 25 amines (10%)7.9 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inula leaves(25%) 12.5 triethanolamines 8.5 Zeofree 80 (95%) Stepsperse DF500 (10%)Monoamine 779 (20%) Triethanolamine (50%) Inula leaves (25%) 25(NH₄)₂HPO₄ (25%) 7.5 Zeofree 80 (25%) diammonium phosphate (25%) ReaxM100 (5%) Kaolin (20%) Inula leaves (25%) 25 (NH₄)₂HPO₄ (25%) 8Hubersorb 600 (25%) diammonium phosphate (25%) Reax M100 (5%) Kaolin(20%) Inula leaves (25%) 25 (NH₄)₂HPO₄(25%) 8 Hubersorb 600 (40%)diammonium phosphate (25%) Stepan DF 500 (10%) Inula leaves (25%) 25(NH₄)₂HPO₄ (25%) 8 Hubersorb 600 (24%) diammonium phosphate (25%) StepanDF 500 (5%) 40% Tdet 09 on Zeolex 7A (5%)

Example 25

[0062] EXAMPLE 25 Late blight in potato. % infected leaf area 6 daysConc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 100 100 100100 Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%)50 25 38 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inulaleaves (25%) 25 25 18 Zeofree 80 (95%) Stepsperse DF500 (10%) Monoamine779 (20%) Triethanolamine (50%) Inula leaves (25%) 100 69 50 Zeofree 80(25%) diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inulaleaves (25%) 100 50 20 Hubersorb 600 (25%) diammonium phosphate (25%)Reax M100 (5%) Kaolin (20%) Inula leaves (25%) 50 20 0 Hubersorb 600(40%) diammonium phosphate (25%) Stepan DF 500 (10%) Inula leaves (25%)100 88 25 Hubersorb 600 (24%) diammonium phosphate (25%) Stepan DF 500(5%) 40% Tdet 09 on Zeolex 7A (5%)

Example 26

[0063] EXAMPLE 26 Late blight in tomato. Suspensions were used ˜100hours after preparation. % infected leaf area 4 days Conc. % a-iFormulation 0 0.25 0.5 1.0 Inula leaves (27%) 95 88 75 38 Zeofree 80(60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%) 19 15 8 Zeofree80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inula leaves (25%) 18 1 0Zeofree 80 (95%) Stepsperse DF500 (10%) Monoamine 779 (20%)Triethanolamine (50%) Inula leaves (25%) 75 63 25 Zeofree 80 (25%)diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves(25%) 75 63 25 Hubersorb 600 (25%) diammonium phosphate (25%) Reax M100(5%) Kaolin (20%) Inula leaves (25%) 50 38 9 Hubersorb 600 (40%)diammonium phosphate (25%) Stepan DF 500 (10%) Inula leaves (25%) 75 3819 Hubersorb 600 (24%) diammonium phosphate (25%) Stepan DF 500 (5%) 40%Tdet 09 on Zeolex 7A (5%)

Example 27

[0064] EXAMPLE 27 Downy mildew in cucumber. % infected leaf area 6 daysConc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 100 63 38 38Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%) 3838 10 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inula leaves(25%) 20 20 13 Zeofree 80 (95%) Stepsperse DF500 (10%) Monoamine 779(20%) Triethanolamine (50%) Inula leaves (25%) 50 30 30 Zeofree 80 (25%)diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves(25%) 44 13 13 Hubersorb 600 (25%) diammonium phosphate (25%) Reax M100(5%) Kaolin (20%) Inula leaves (25%) 44 8 8 Hubersorb 600 (40%)diammonium phosphate (25%) Stepan DF 500 (10%) Inula leaves (25%) 75 6344 Hubersorb 600 (24%) diammonium phosphate (25%) Stepan DF 500 (5%) 40%Tdet 09 on Zeolex 7A (5%)

Example 28

[0065] EXAMPLE 28 Downy mildew in grapes. % sporulating leaf area 8 daysConc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 97 60 5 0Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%) 2525 20 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inula leaves(25%) 50 25 15 Zeofree 80 (95%) Stepsperse DF500 (10%) Monoamine 779(20%) Triethanolamine (50%) Inula leaves (25%) 70 15 0 Zeofree 80 (25%)diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves(25%) 60 20 20 Hubersorb 600 (25%) diammonium phosphate (25%) Reax M100(5%) Kaolin (20%) Inula leaves (25%) 50 25 20 Hubersorb 600 (40%)diammonium phosphate (25%) Stepan DF 500 (10%) Inula leaves (25%) 80 8050 Hubersorb 600 (24%) diammonium phosphate (25%) Stepan DF 500 (5%) 40%Tdet 09 on Zeolex 7A (5%)

Example 29

[0066] EXAMPLE 29 Powdery mildew in wheat. % infected leaf area 8 daysConc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 50 50 25 13Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%) 2513 0 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inula leaves(25%) 25 13 0 Zeofree 80 (95%) Stepsperse DF500 (10%) Monoamine 779(20%) Triethanolamine (50%) Inula leaves (25%) 38 19 13 Zeofree 80 (25%)diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves(25%) 38 25 13 Hubersorb 600 (25%) diammonium phosphate (25%) Reax M100(5%) Kaolin (20%) Inula leaves (25%) 30 25 0 Hubersorb 600 (40%)diammonium phosphate (25%) Stepan DF 500 (10%) Inula leaves (25%) 25 255 Hubersorb 600 (24%) diammonium phosphate (25%) Stepan DF 500 (5%) 40%Tdet 09 on Zeolex 7A (5%)

[0067] In view of the teachings presented herein, other modificationsand variations of the present inventions will be readily apparent tothose of skill in the art. The foregoing discussion, and description areillustrative of some embodiments of the present invention, but are notmeant to be limitations on the practice thereof. It is the followingclaims, including all equivalents, which define the scope of theinvention.

[0068] Any patents or publications mentioned in this specification areindicative of the levels of those skilled in the art to which theinvention pertains. These patents and publications are hereinincorporated by reference to the same extent as if each individualpublication was specifically and individually indicated to beincorporated by reference.

[0069] One skilled in the art will readily appreciate that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those inherent therein. The presentexamples along with the methods, procedures, treatments, and specificcompositions described herein are presently representative of preferredembodiments, are exemplary, and are not intended as limitations on thescope of the invention. Changes therein and other uses will occur tothose skilled in the art which are encompassed within the spirit of theinvention as defined by the scope of the claims.

What is claimed:
 1. A method for preparing an extract from Inulaspecies, said method comprising the steps of: (a) contacting shootsand/or portions thereof with an alkaline aqueous solvent to form asolution and debris; and (b) removing the debris from the solution.
 2. Amethod according to claim 1, wherein said removing step includesfiltering the debris from the solution.
 3. A method according to claim1, wherein the shoots include leaves and stems of upper parts of theshoots.
 4. A method according to claim 1, wherein the shoots or portionsthereof are dried prior to said contacting step.
 5. A method accordingto claim 4 including the step of grinding the dried shoots or portionsthereof to form a powder.
 6. A method according to claim 1, wherein theshoots or portions thereof are freshly cut.
 7. A method according toclaim 1, wherein the alkaline aqueous solvent comprises at least oneamino compound.
 8. A method according to claim 7, wherein the aminocompound comprises an ammonium compound.
 9. A method according to claim7, wherein the amino compound is selected from the group consistingessentially of organic amines and heterocyclic amines.
 10. A methodaccording to claim 9, wherein the organic amine compound is selectedfrom the group consisting essentially of ethylamine, ethylenediamine,diaminopropane, diethanolamine, triethanolamine, and butylamine.
 11. Amethod according to claim 9, wherein the heterocyclic amine compound isselected from the group consisting essentially of piperazine andpiperidine.
 12. A method according to claim 7, wherein the alkalineaqueous solvent comprises inorganic bases or salts thereof.
 13. A methodfor protecting plants against fungal infection, said method comprisingthe steps of: (a) preparing a fungicidal extract of Inula species bycontacting shoots and/or portions thereof with an alkaline aqueoussolvent to form an extract solution and debris; and (b) applying afungicidally effective amount of the fungicidal extract to a plant forprotecting the plant against fungal infection.
 14. A method according toclaim 13, wherein the plant includes grapevines, cucurbits, tomatoes,wheat, barley, onion, tobacco, crucifers, and potatoes.
 15. A methodaccording to claim 13, wherein the fungal infection is caused byphytopathogenic fungi including Oomycetes, Ascomycetes, Basidiomycetesand Fungi imperfecti classes.
 16. A method according to claim 15,wherein the fungal infection is caused by a fungus includingCladosporium cucumerinum, Phytophthora infestans, Botrytis cinerea,Pseudoperonospora cubensis, Sphaerotheca fuliginea, Plasmopara viticola,Uromyces appendiculatis, and Erysiphe graminis.
 17. A method accordingto claim 13, wherein said preparing step includes filtering the debrisfrom the solution.
 18. A method according to claim 13, wherein theshoots include leaves and stems of upper parts of the shoots.
 19. Amethod according to claim 13, wherein the shoots or portions thereof aredried prior to said contacting step.
 20. A method according to claim 19including the step of grinding the dried shoots or portions thereof toform a powder.
 21. A method according to claim 13, wherein the shoots orportions thereof are freshly cut.
 22. A method according to claim 13,wherein the alkaline aqueous solvent comprises at least one aminocompound.
 23. A method according to claim 22, wherein the amino compoundcomprises an ammonium compound.
 24. A method according to claim 22,wherein the amino compound is selected from the group consistingessentially of organic amines and heterocyclic amines.
 25. A methodaccording to claim 24, wherein the organic amine compound is selectedfrom the group consisting essentially of ethylamine, ethylenediamine,diaminopropane, diethanolamine, triethanolamine, and butylamine.
 26. Amethod according to claim 24, wherein the heterocyclic amine compound isselected from the group consisting essentially of piperazine andpiperidine.
 27. A method according to claim 13, wherein the alkalineaqueous solvent comprises inorganic bases or salts thereof.
 28. A methodfor preparing a fungicide derived from Inula species, said methodcomprising the step of: combining a substantially powdered form of Inulashoots and/or portions thereof with a solid chemical to form a mixturewhich upon dissolution of the mixture in an aqueous solution, forms analkaline aqueous solution which can be directly applied to plants toprevent fungal infection.
 29. A method according to claim 28, whereinthe solid chemical comprises at least one amino compound.
 30. A methodaccording to claim 29, wherein the amino compound comprises an ammoniumcompound.
 31. A method according to claim 29, wherein the amino compoundis selected from the group consisting essentially of organic amines andheterocyclic amines.
 32. A method according to claim 31, wherein theorganic amine compound is selected from the group consisting essentiallyof ethylamine, ethylenediamine, diaminopropane, diethanolamine,triethanolamine, and butylamine.
 33. A method according to claim 31,wherein the heterocyclic compound is selected from the group consistingessentially of piperazine and piperdine.
 34. A method according to claim28, wherein the solid chemical comprises inorganic bases or saltsthereof.
 35. A method according to claim 28, wherein an emulsifier isadded to the mixture.
 36. A method according to claim 35, wherein theemulsifier comprises a surfactant.
 37. A method according to claim 36,wherein the surfactant comprises sodium dodecyl sulfate.
 38. A methodaccording to claim 35, wherein the emulsifier comprises diatomaceousearth.